Effect of tetrandrine on cytokines in LPS-stimulated RAW264.7 cell

Objective To investigate the effects of tetrandrine(Tet) on expressions of proinflammatory and anti-inflammatory cytokine in lipopolysaccharide(LPS)-stimulated RAW264.7 cells.Methods RAW 264.7 cells were cultured in media containing 1 μg/mL LPS and different doses of Tet.The proliferation of cells was examined by MTT assay,and nuclear translocation of NF-κB was detected and analyzed by the laser scanning confocal microscope(LSCM),and the levels of IL-6,TNF-α,and IL-10 were tested by ELISA.Results Compared with the control group,Tet had no influence on the proliferation of RAW264.7 cells no matter with LPS or not when its concentration lower than 1 μmol/L;But when its concentration higher than 10 μmol/L,Tet significantly inhibited the growth of the cells whether LPS existed or not.Meanwhile,Tet significantly decreased the levels of IL-6,and TNF-α induced by LPS,and upregulated IL-10 in supernatant.At the mean time,results from LSCM showed that in Tet 1 μmol/L group,positive cells(dyeing-red p65 of NF-κB was translocated into nucleuses and made the nuclueses red) decreased siglificantly,while negative cells(dyeing-red p65 of NF-κB was collectted in endochylema and nucleuses became vacant) were predominantly founded.Conclusion Tet has anti-inflammatory effect,which might be mediated by downregulating inflammatory factors IL-6,TNF-α through NF-κB signal pathway as well as increasing levels of anti-inflammatory factor IL-10.